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. Author manuscript; available in PMC: 2020 Apr 17.
Published in final edited form as: J Am Chem Soc. 2019 Apr 8;141(15):6204–6212. doi: 10.1021/jacs.8b12954

Figure 2.

Figure 2.

CRACR mediated protein labeling is orthogonal to SPAAC and SPIEDAC. A) Treatment of three sfGFP mutants (incorporating 5HTP, pAzF or CpK at the 151 position) with 4CDZ, DBCO-tamra, and tetrazine-fluorescein followed by ESI-MS analysis reveals selective labeling of 5HTP, pAzF, and CpK by CRACR, SPAAC, and SPIEDAC, respectively. B) Analysis of protein samples from a similar experiment (4CDZ-biotin was used instead of 4CDZ) by SDS-PAGE followed by fluorescence imaging (anti-biotin Western blot for proteins treated with 4CDZ-biotin) further confirms selective labeling of each mutant with the intended reagent.